The real-time Delphi method represents a refinement of the classical Delphi technique, designed to overcome limitations such as prolonged study duration and delayed feedback during consensus development. This article, building upon the classical Delphi foundation, systematically elaborates on the application process, advantages, and limitations of the real-time Delphi method. It further presents currently available websites or software capable of facilitating real-time Delphi exercises and offers considerations and recommendations for its application in guideline development, aiming to serve as a reference for relevant researchers.

Objective:To search for potential synergistic drugs that can enhance the inhibitory effect of lenvatinib on the proliferation of hepatocellular carcinoma cells by compound screening and explore the underlying molecular mechanisms.Methods:The impact of lenvatinib on the proliferation of Huh7 cells was detected using CCK-8 assay and long-term cell proliferation assays.The potential synergistic drugs for lenvatinib in Huh7 cells was screened using an FDA-approved compound library.Subsequently,the inhibitory effect of lenvatinib combined with zinc pyrithione on the proliferation of Huh7 cells was assessed using CCK-8 assay and long-term cell proliferation assay.Furthermore,RNA-sequencing was utilized to investigate the changes in gene expression profiles following the combined action of zinc pyrithione and lenvatinib,exploring the potential molecular mechanisms.The impact of combination therapy on signaling pathways was investigated through Gene Set Enrichment Analysis(GSEA)and Gene Ontology Analysis(GO).Results:Lenvatinib exerts dose-dependent inhibition on the proliferation of Huh7 in vitro(IC50 value=0.190 μmol/L).Screening of compound libraries identified zinc pyrithione as a compound that synergistically promotes the inhibition of Huh7 cell proliferation by lenvatinib.This result was further validated through CCK-8 assay(P<0.05)and long-term cell proliferation experiments.Compared to treatment with lenvatinib alone,the combination treatment of zinc pyrithione and lenvatinib upregulated signaling pathways related to oxidative stress response,apoptosis,and cellular responses to copper ions in Huh7 cells,with Gene Set Enrichment Analysis(GSEA)showing significant enrichment of the oxidative phosphorylation pathway.Conclusion:Lenvatinib can inhibit the proliferation of Huh7 cells in vitro,while Zinc Pyrithione can synergize with lenvatinib to exert a more significant inhibitory effect on the Huh7 cells.The molecular mechanism of this synergistic effect may involve generation of a large amount of reactive oxygen species(ROS)to induce apoptosis of liver cancer cells through oxidative stress response,as well as promoting the death of hepatocellular carcinoma cells by disrupting copper homeostasis.

Objective This study aims to investigate the role of gap junction mediated by connexin 43(Cx43)in renal injury induced by periodontitis in rats.Methods Twelve SPF-grade Wistar male rats were divided into a control group and a periodontitis group by using a completely random number table method,with six rats in each group.The control group rats were not treated,while the periodontitis group rats were subjected to wire ligation of the neck of their bilateral maxillary first molars to construct a periodontitis model.After 8 weeks of modeling,the rats were examined for clinical indicators of the periodontium.micro-CT scanning of the maxilla reconstructed its 3D structure and analyzed the absorption of alveolar bone.Histopathological changes in periodontal and renal tissues were detected.MitoSOX red reagent was used to determine reactive oxygen species(ROS)content in renal tissues.A biochemical reagent kit was used to detect serum oxidative stress biomarkers.Real-time fluo-rescent quantitative-polymerase chain reaction(qRT-PCR)was employed to determine Cx43,nuclear factor kappa-B(NF-κB),interleukin(IL)-1β,IL-6,BCL2-Associated X(Bax),B-lymphomatoma-2 gene(Bcl-2),and Caspase-3 mRNA were determined.Western blot analysis was used to detect Cx43,NF-κB,IL-1β,Bax,Bcl-2 and Caspase-3 protein.Re-sults micro-CT 3D reconstruction showed significant bone resorption of the first molar alveolar bone in the periodonti-tis group rats and decreased height of the alveolar ridge.The distance from the enamel cementum boundary to the top of the alveolar ridge in the periodontitis group was significantly higher than that inthe control group.The histopathological results showed a large number of inflammatory cells that infiltrated the periodontal tissue of the periodontitis group,and the alveolar bone was significantly absorbed.Rats in the periodontitis group also exhibited mild thickening of the glomer-ular basement membrane,dilation of the Bowman's capsule,and destruction of the brush-like edge of the renal tubules in the renal tissue.The MitoSOX red staining results showed a significant increase in ROS content in the renal tissue of the periodontitis group.The biochemical test results showed that the levels of superoxide dismutase and glutathione in the serum of rats with periodontitis decreased,while that of malondialdehyde increased.The results of qRT-PCR and Western blot showed that the expression levels of Cx43,IL-1β,IL-6,Bax,Caspase-3 mRNA and Cx43,IL-1β,NF-κB,Bax,Caspase-3 proteins in the periodontitis group significantly increased compared with those in the control group,while the expression levels of Bcl-2 mRNA and protein decreased.Conclusion Periodontitis may activate NF-κB sig-naling molecules by upregulating the expression of Cx43 in rat kidney tissues,leading to increased levels of inflamma-tion and apoptosis and ultimately inducing kidney injury.

Objective:To search for potential synergistic drugs that can enhance the inhibitory effect of lenvatinib on the proliferation of hepatocellular carcinoma cells by compound screening and explore the underlying molecular mechanisms.Methods:The impact of lenvatinib on the proliferation of Huh7 cells was detected using CCK-8 assay and long-term cell proliferation assays.The potential synergistic drugs for lenvatinib in Huh7 cells was screened using an FDA-approved compound library.Subsequently,the inhibitory effect of lenvatinib combined with zinc pyrithione on the proliferation of Huh7 cells was assessed using CCK-8 assay and long-term cell proliferation assay.Furthermore,RNA-sequencing was utilized to investigate the changes in gene expression profiles following the combined action of zinc pyrithione and lenvatinib,exploring the potential molecular mechanisms.The impact of combination therapy on signaling pathways was investigated through Gene Set Enrichment Analysis(GSEA)and Gene Ontology Analysis(GO).Results:Lenvatinib exerts dose-dependent inhibition on the proliferation of Huh7 in vitro(IC50 value=0.190 μmol/L).Screening of compound libraries identified zinc pyrithione as a compound that synergistically promotes the inhibition of Huh7 cell proliferation by lenvatinib.This result was further validated through CCK-8 assay(P<0.05)and long-term cell proliferation experiments.Compared to treatment with lenvatinib alone,the combination treatment of zinc pyrithione and lenvatinib upregulated signaling pathways related to oxidative stress response,apoptosis,and cellular responses to copper ions in Huh7 cells,with Gene Set Enrichment Analysis(GSEA)showing significant enrichment of the oxidative phosphorylation pathway.Conclusion:Lenvatinib can inhibit the proliferation of Huh7 cells in vitro,while Zinc Pyrithione can synergize with lenvatinib to exert a more significant inhibitory effect on the Huh7 cells.The molecular mechanism of this synergistic effect may involve generation of a large amount of reactive oxygen species(ROS)to induce apoptosis of liver cancer cells through oxidative stress response,as well as promoting the death of hepatocellular carcinoma cells by disrupting copper homeostasis.

Objective:To investigate Helicobactor pylori (H. pylori) infection status and interfamilial transmission pattern in Zhengzhou area. Methods:A cross-sectional study was conducted from September 2020 to march 2021, among 731 individual from 266 families randomly selected from 9 communities of Zhengzhou area. H. pylori infection status was determined by serum antibody tests, and 13C-urea breath test was performed in the previously eradicated population to clarify the current infection status. The individual and familial infection rate, infection status for couples and children and adolescent were analyzed. Results:Among 731 individuals from 266 families, 397 of them were H. pylori positive. The individual infection rate was 54.31% (397/731); among infected individuals 77.83% (307/397) were infected with type Ⅰ strain, 22.67% (90/397) were infected by type Ⅱ strain. Annual household income ( χ2=0.419, 0.410, 0.213, all P>0.05), smoking history (χ 2=0.071, P>0.05), drinking history ( χ2=0.071, P>0.05), dining place ( χ2=0.009, P>0.05), gastrointestinal symptoms ( χ2=0.047, P>0.05), family history of gastric disease ( χ2=0.069, P>0.05), and history of gastric cancer ( χ2=0.004, P>0.05) had no significant differences between H. pylori-positive and -negative groups, but the infection rate in individuals with higher education level was lower ( χ2=4.449, P<0.05). The infection rate was significantly higher in≥18 age groups compared with<18 age groups ( χ2=6.531, 23.362, 20.671, 24.244, 37.948, 14.597 and 5.170, all P<0.05). The familial H. pylori infection rate was 87.59% (233/266), and in 61 families all member were infected (26.18%, 61/233). The positive rate was 23.08% (6/26) in 50 families with children under 18 years when both parents were infected. Among 231 coupled families, both couples were infected in 78 families (33.76%), one couple was infected in 113 families (48.92%), and both couples were not infected in 40 (17.32%). With the increase of marriage time, the infection rate of both spouses increased significantly ( χ2=7.775, 12.662, 15.487, all P<0.05). Conclusions:The distribution of H. pylori infection presents a family cluster pattern, and intrafamilial infection is an important transmission rout of H. pylori. The type I strain of H. pylori is the dominate strain in this area.

Objective:To analyze the epidemiological characteristics of imported COVID-19 cases in early phase in Shanghai, introduce measures and provide reference for prevention and control of imported COVID-19 cases.Methods:Data of imported COVID-19 cases in Shanghai reported as of 30 March, 2020 were obtained from National Notifiable Disease Report System of China CDC and field epidemiological investigation reports by CDCs in Shanghai. The information about measures of prevention and control was collected from official websites and platforms of the governments. Data cleaning and statistical analysis were performed with softwares of EpiData 3.1, Excel 2019 and SAS 9.4.Results:A total of 171 imported COVID-19 cases had been reported as of 30 March, 2020 in Shanghai, including 170 confirmed cases and 1 asymptomatic infection case. Among them, cases of Chinese nationality accounted for 71.3% (122/171) and cases of foreign nationality accounted for 28.7% (49/171). The median age of the cases was 23 years ( P 25, P 75: 18, 35 years), and the male to female ratio of the cases was 1.3∶1. Students accounted for 56.6% (97/171). About 45.6% (78/171) of the cases fell ill before arriving in Shanghai. The cases with mild or common clinical manifestation accounted for 96.5% (165/171) and no significant difference in clinical type was observed between overseas Chinese cases and foreign cases. The epidemic curve by diagnosis date reached peak on March 24, and the number of the cases gradually declined due to the closed-loop management process of joint port prevention and control mechanism. The 171 imported COVID-19 cases were mainly from 24 countries and regions, including the United Kingdom (64 cases, 37.3%), the United States (32 cases, 18.6%), France (19 cases, 11.0%) and Italy (16 cases, 9.4%). About 40.4% of the cases (69/171) planned to continue travelling to 21 other provinces and municipalities in China. Customs quarantine and community observation/detection points identified 43.9% (75/171) cases and 31.0% (53/171) cases, respectively. Conclusions:The imported COVID-19 cases in early phase in Shanghai were mainly young population and students accounted for high proportion. The imported risk of COVID-19 was consistent with the severity of the epidemic in foreign countries. The closed-loop management model of the joint port prevention and control mechanism plays an important role in the identification and management of the imported COVID-19 cases.

Objective To investigate the differences in tumor volume and metastatic tumors of the liver and regional lymph nodes between contrast-enhanced computed tomography (CT) and diffusion-weighted magnetic resonance imaging (DWMRI) through a comparative analysis,as well the useful information for target volume delineation,and to guide radiotherapy in clinical practice.Methods A total of 40 patients with pancreatic cancer were enrolled and underwent contrast-enhanced CT and DWMRI in the same position.The target volume was delineated,the major axis of the maximum tumor section was measured,and the numbers of liver metastatic tumors and metastatic tumors of the lymph nodes with a diameter of 5-8 mm or＞8 mm were measured based on the CT and DWMRI images.The analysis was performed by using paired t-test or paired Wilcoxon rank sum test.Results The mean gross tumor volume (GTV) delineated by contrast-enhanced CT and DWMRI was 54.95 cm3 and 41.67 cm3(P =0.000),and the mean value-different value was 13.28 cm3.The major axis of the maximum tumor section measured by contrast-enhanced CT and DWMRI were 4.18 cm and 3.94 cm (P=0.000),respectively,and in two patients,dCT was smaller than dDWMRI.A total of 83 liver metastatic tumors were identified by contrast-enhanced CT,and 112 were identified by DWMRI;the liver metastatic tumors detected by contrast-enhanced CT accounted for 74％ of those detected by DWMRI.As for the metastatic tumors of the lymph nodes with a diameter of 5-8 mm or＞8 mm,103 or 46 were detected by contrast-enhanced CT,and 200 or 56 were detected by DWMRI,and the tumors detected contrast-enhanced CT accounted for 52％ or 82％ of those detected by DWMRI.There were significant differences in all data between contrast-enhanced CT and DWMRI.Conclusions GTV and the major axis of the maximum tumor section measured by DWMRI are lower than those measured by contrast-enhanced CT,and contrast-enhanced CT is sensitive in detecting the metastatic tumors of the liver and lymph nodes.However,it is necessary to conduct further controlled experiments with reference to pathology.

Objective To observe the clinical efficacy of Jianpi Huatan Decoction plus Zhongwan (RN12) moxibustion on patients of mild to moderate stable chronic obstructive pulmonary disease (COPD) with syndrome of qi deficiency of lung and spleen, and analyze the related influencing factors. Methods With prospective randomized clinical trial, 140 cases of mild to moderate stable COPD were randomized into two groups (70 cases for each group). Treatment group was treated by Jianpi Huatan Decoction plus Zhongwan moxibustion based on western medicine, and control group was only treated by western medicine. Both groups were treated for 12 weeks and followed up for 12 weeks. Scores of clinical symptoms and quality of life questionnaire were examined, and multiple logistic regression analysis was used to find possible impacting factors on the effect. Results Symptoms were significantly alleviated after treatment (P<0.01) in both groups. The clinical effects and quality of life in treatment group were significantly improved than the control group (P<0.01). There were no significant changes in life quality of control group ( P>0.05). Multiple logistic regression analysis showed that different therapies and severity of COPD were the major influencing factors of clinical effect (P<0.05, P<0.01). Conclusion Jianpi Huatan Decoction plus Zhongwan moxibustion had significant therapeutic effect on mild to moderate stable COPD patients with syndrome of qi deficiency of lung and spleen by alleviating symptoms and improving quality of life. Treatment style and severity of COPD are the main influencing factors.

Objective To investigate the changes of serum soluble factor-related apoptosis (sFas) and soluble Fas ligand (sFasL) and their relations with cognition disorders in the patients with vascular dementia (VaD). Methods Serum concentrations of sFas and sFasl were detected by enzyme-linked immunosorbent assay (ELISA) and compared between 70 patients with VaD aged (72.5± 7.5)years and 50 healthy elderly people aged(72.5 ± 7.5)years.The VaD patient's cognitive functions were evaluated by activity of daily living scale (ADL),mini mental state examination (MMSE) and hachinski ischemia score (Hachinski). Results The serum levels of sFas and sFasL in VaD patients were (228.0±60.7)μg/L and (146.8±30.1)μg/L,and in the healthy elderly were (62.4±22.6)μg/L and (82.3 ± 18.7)μg/L,respectively.The serum levels of apoptosis factors in VaD patients were significantly higher than in the healthy controls (t=20.883,14.453,P＜0.01).sFas level was negatively correlated with age,the scores of ADL and Hachiuski while positively with the scores of MMSE (r=-0.956,-0.943,-0.950 and 0.904,all P＜0.01). sFasL level was negatively correlated with the scores of MMSE while positively with age,the scores of ADL and Hachinski (r=-0.899,0.963,0.948 and 0.939,a11 P＜0.01). Conclusions Apoptosis may be involved in the pathological change during VaD and the serum levels of sFas and sFasL might be related with cognition disorders.

Objective To explore the method for establish the temporal lobe epilepsy model in SD rats with stereotactic technique and to study the characteristic of electroencephalography (EEG) after epileptic seizure. Methods Kainic acid was injected into right hippocampus via stereotactic technique in 20 rats, and the behavior of rat was recorded by a video camera. The electrical activity changes of brain were recorded by EEG in bilateral hippocampus, amygdala and frontal lobe. Results According to Racine's scale, 6 rats were typeⅤ,10 rats were type Ⅳ,3 rats were type Ⅲ,1 rat was type Ⅱ. The spike wave was originated from hippocampus and spread to amygdala and frontal lobe. Pathologic changes showed that prolonged seizures induced pyramidal cell loss and death in hippocampus. Conclusions The method to establish kainic acid induced temporal lobe epilepsy model with stereotactic technique was credible, economical and useful. The spike wave originated from hippocampus was the basic feature of EEG of temporal lobe epilepsy model in rats.